Phytochemical Characterization of Cannabis sativa L. Roots from Northeastern Brazil

This article describes the phytochemical study of Cannabis sativa roots from northeastern Brazil. The dried plant material was pulverized and subjected to exhaustive maceration with ethanol at room temperature, obtaining the crude ethanolic extract (Cs-EEBR). The volatile compounds were analyzed by gas chromatography coupled with mass spectrometry (GC/MS), which allowed to identify 22 compounds by comparing the linear retention index (LRI), the similarity index (SI) and the fragmentation pattern of the constituents with the literature. By this technique the major compounds identified were: friedelan-3-one and β-sitosterol. In addition, two fractions were obtained from Cs-EEBR by classical column chromatography and preparative thin layer chromatography. These fractions were analyzed by NMR and IR and together with the mass spectrometry data allowed to identify the compounds: epifriedelanol, friedelan-3-one, β-sitosterol and stigmasterol. The study contributed to the phytochemical knowledge of Cannabis sativa, specifically the roots, as there are few reports on the chemical constituents of this part of the plant.     

Separation of Soybean Sterols by Florisil Chromatography and Characterization of Acylated Steryl Glucoside

Florisil column chromatography was demonstrated to be effective in differentiation between different forms of sterols. Sterols of ground soybeans are in four forms, free, ester, and free and acylated glucosides, as analyzed on acetone extracts. In soybean oil foots, steryl ester is present in negligibly small amount. The acylated steryl glucosides were isolated from oil foots in a crystalline state. A chemical structure, steryl 6-acyl d-glucoside, was assigned to the compound, and its probable identity with the glucosides reported by Lepage is discussed. The acylated glucoside preparation was shown to be heterogeneous in composition, carrying palmitic, stearic, oleic, linoleic and linolenic acids as the main acyl moieties and campesterol, stigmasterol and β-sitosterol as steryl moieties. The presence of the three sterols is common to three other forms of sterols.     

An NAD(P)H Model Reaction: Reduction of α-Keto Esters by the Hantzsch Ester Accelerated by Zinc Complex in the Reformatsky Mixture

Aiming to get useful steroidal alkaloids by tissue culture of Solanum laciniatum Ait., indefinitely growing callus tissue was prepared from the mother plant. Some nutritional requirements for the growth of the callus tissue were studied. By examining steroidal compounds in callus culture, cholesterol, stigmasterol, β-sitosterol, lanosterol, squalene, diosgenin and a new steroidal alkaloid were found to be formed in the callus culture. The new steroidal alkaloid was found to be solasodine derivative containing rhamnose and other unidentified sugars.     

Sterols in Germinating Seeds and Developing seedlings of Longleaf Pine, Pinus palustris

Sterols in germinating embryos and young seedlings of longleaf pine (Pinus palustris Mill.) were identified and quantities determined for different periods after germination. Sterol analyses were performed by gas-liquid chromatography (GLC) and verified by combination of GLC-mass spectrometry. Campesterol and β-sitosterol were two major sterols which accounted for most of the sterol composition while stigmasterol was present in very small amounts. No cholesterol was revealed by GLC-mass spectrometry although there was a minor peak appearing on the sterol gas-liquid chromatograms with a retention time close to that of authentic cholesterol. By fractionation, three different forms of sterols were obtained: steryl esters, steryl glycosides, and free sterols. The sterols were mainly found in the esterified fraction, while steryl glycosides and free sterols only made up a small portion of the total sterol value. The total sterol content in general increased during seedling development, and this increase reflected mainly a change in steryl esters. The low levels of both free and glycosidic sterols remained nearly unchanged throughout the experimental germination period.     

Studies on Fatty Acids and Unsaponifiable Constituents of Oil from Tubers of Cyperus esculentus L.

The fatty acids of the oil from tubers of Cyperus esculentus L. were determined by gas chromatography with DC-11 and DEGS stationary phases. Oleic, linoleic, palmitic and stearic acids are the major constituents in the fatty acid fraction, while lauric, myristic, linolenic, arachidic, dadoleic, behenic and tetracosanoic acids are the minor ones. The unsaponifiable matters of the oil were separated by column chromatography with silica gel and thin layer chromatography with silica gel G into three fractions: sterols, 4-methylsterols and triterpene alcohols. The acetates of sterols, 4-methylsterols and triterpene alcohols were separated by TLC with 20% silver nitrate impregnated silica gel G, using CH2Cl2-petroleum system as developing reagents. The identification of major components was carried out by TLC, mp, optical rotation, GLC, IR spectrum and GC-MS. It was found that β-sitosterol, stigmasterol and campesterol were present in large amounts in the unsaponifiable fraction, β-sitosterol, stigmasterol, △5-and △7-avenasterol, 24- methylenecholesterol and 24-methylenecholest-7-enol in the sterol fraction, obtusifoliol, gramisterol and citrostadienol in the 4-methylsterol fraction, and cycloartanol, cydoartenol, 24- methylenecydoartanol and cyclobranol in the triterpene alcohol fraction were isolated and identified, while campesterol, campestanol, stigmastanol, △7-stigmastenol, △7-campestenol and △7-cholestenol were identified only. We found no evidence of the occurence of nonedibles in this oil.     

拳卷地钱脂溶性部分化学成分研究

为了研究拳卷地钱脂溶性部分化学成分 ,采用硅胶色谱柱分离 ,物理、化学和光谱方法鉴定结构 ,分离并鉴定了β 谷甾醇和豆甾醇 2个化合物 ,均为首次从该植物中分离得到。     

Intake of stigmasterol and β-sitosterol alters lipid metabolism and alleviates NAFLD in mice fed a high-fat western-style diet

Objective

To investigate and compare the effects of two common dietary phytosterols, stigmasterol and β-sitosterol, in altering lipid metabolism and attenuating nonalcoholic fatty liver disease (NAFLD).

Pollen size evolution: correlation between pollen volume and pistil length in Asteraceae

Based on the assumptions that pollen tube length is predetermined by provisions in the pollen and that it is a function of pistil length, I hypothesise that species with longer pistils will have larger pollen grains than species with shorter pistils, and that pistil length and pollen size will be positively correlated in a linear manner. To test this hypothesis, the relationship between pollen grain volume and pistil length was compared in 43 Asteraceae species from Argentina. A positive linear correlation was found between pollen volume and pistil length. This correlation remained significant even after potential effects of phylogenetic relatedness were removed. The maintenance of this correlation suggests that in Asteraceae the association between pistil length and pollen volume may reflect a functional rather than a phyletic relationship. In addition, the pistil length: pollen volume ratio (PPR) was analysed in relation to the phylogenetic position of the species. High values of PPR would imply a reduction of the male gametophyte in relation to the minimal volume that a pollen grain must have to grow and fertilise an ovule. Thus, the general pattern of pollen volume reduction in relation to pistil length previously found among many angiosperm families will be also present within a family, i.e., PPR values of derived Asteraceae would be higher than those of basal species. Results indicated that reduction of pollen volume in derived Asteraceae was three times greater than the concomitant shortening of pistil length. Consequently, PPR increased with the phylogenetic position of the taxa. This work supports the correlation between pistil and pollen characters previously found for other plant families and confirms the influence of post-pollination processes on pollen size evolution. Received: 4 November 1999 / Revision accepted: 14 February 2000     

Dietary Sterol Preference in the Silkworm,Bombyx mori

Since insects are unable to biosynthesize sterols de novo, sterols must be obtained from dietary sources. Although it has been reported that β-sitosterol is crucial for larval growth in the silkworm, Bombyx mori, little has been investigated concerning the dietary selection of sterols by Bombyx larvae. Here, we demonstrate that Bombyx larvae have the following sterol preference: β-sitosterol >> ergosterol > cholesterol = stigmasterol. Interestingly, Bombyx larvae preferred ergosterol, an inhibitory sterol on larval growth, indicating that sterol selection following first contact of the diet with the mouthpart might be different from the sterol recognition mechanism present in sterol metabolism.     

Changes in Sterol Composition during Greening of Etiolated Barley Shoots

The following sterols were identified in barley shoots: stigmasterol, β-sitosterol, campesterol, and cholesterol. The total sterol content of green and etiolated tissue was 2.84 and 3.20 milligrams per gram dry weight, respectively. The free sterols accounted for most of the difference in total sterol content. The sterol ester, sterol glycoside, and acylated sterol glycoside contents of green and etiolated barley shoots were essentially the same. Etiolated tissue had twice as much total β-sitosterol as stigmasterol, while green tissue had equal amounts of these two sterols. The campesterol and cholesterol content was the same in green and etiolated tissue. This same sterol composition pattern held true for the free, glycosidic, and acylated glycosidic sterols; however, the sterol ester fraction had a completely different composition pattern. The esterified stigmasterol content was quite low in green and etiolated tissue, and campesterol was the second largest esterfied sterol component in etiolated tissue. Etiolated barley seedlings exposed to light had a shift in the ratio of free stigmasterol to β-sitosterol in favor of stigmasterol; however, no correlation was observed between chlorophyll synthesis and shift in sterol composition.     

Volubilin,a new isoflavone-C-glycoside from Dalbergia volubilis flowers

Dalbergia volubilis flowers after successive extraction with light petroleum, benzene and alcohol yielded β-sitosterol, stigmasterol, biochanin-A and a new isoflavone-C-glycoside named volubilin. The glvcoside on boiling with HI and phenol gave genistein and on ferric chloride oxidation rhamnose. The compound was identified as 8-C-rhamnopyranosyl-genistein-4′,7-dimethyl ether by means of chemical reactions as well as from spectral data.     

Plant sterol-enriched soy milk consumption modulates 5-lipoxygenase, 12-lipoxygenase,and myeloperoxidase activities in healthy adults – a randomized-controlled trial

A randomized, double blind, placebo-controlled and crossover study was conducted to simultaneously measure the effects, after 3-h and 4-week daily exposure to plant sterols-enriched food product, on inflammation, oxidative status, 5-lipoxygenase, 12-lipoxygenase, and myeloperoxidase activities in healthy adults. Eighteen healthy participants (67% female, 35.3 (mean)?±?9.5 (SD) years, mean body mass index 22.8?kg?m^?2) received two soy milk (20g) treatments daily: placebo and one containing 2.0?g free plant sterols equivalent of their palmitates (β-sitosterol, 55%; campesterol, 29%; stigmasterol, 23%). F₂-isoprostanes, leukotriene B₄, sitosterol, campesterol, and stigmasterol concentrations were measured in the blood plasma and urine, using stable isotope-labeled gas chromatography–mass spectrometry. High-sensitivity c-reactive protein, tumor necrosis factor-_α, and lipoxin A₄ concentrations in blood serum were measured using commercially available enzyme immunoassays. Myeloperoxidase activity, serum lipid hydroperoxides, plasma and urinary F₂-isoprostanes, plasma and urinary leukotriene B₄, and plasma high-sensitivity c-reactive protein concentrations were significantly reduced, while circulating lipoxin A₄ concentrations were significantly elevated after 4-week plant sterols treatment. Plant sterols treatment decreased plasma leukotriene B₄ and increased plasma lipoxin A₄ concentrations acutely. Total plant sterols, β-sitosterol, campesterol, and stigmasterol concentrations were significantly elevated after 4-week treatments compared with the pre-treatment concentrations. Our results suggest that dietary plant sterols, in the combination used, can alleviate lipid peroxidation and inflammatory events in vivo. These effects are possibly exerted via the modulation of myeloperoxidase, 5-lipoxygenase, and 12-lipoxygenase activities.     

迷果芹根化学成分的研究

利用普通硅胶柱色谱技术和多种现代波谱技术,从迷果芹(Sphallerocarpus gracilis)的根中分离并鉴定了11个化合物,通过波谱学方法鉴定为:falcarinol(1)、(3R,8S)-falcarindiol(2)、diplaniol(3)、5-hydroxy-8-methoxy-2H-1-benzopyran-2-one(4)、esculetin(5)、东莨菪内酯(6)、xanthalin(7)、isoimperatorin(8)、胡萝卜苷(9)、β-谷甾醇(10)和豆甾醇(11),其中化合物1～6为首次从该植物中分离得到。     

Effects of temperature and gibberellic acid on 4-desmethylsterol composition of Avena sativa stem segments

Of the three morphological subunits of Avena sativa stem segments (node, leaf-sheath and internode) examined, internodes constituted the richest source of phospholipids and sterols, yielding almost double the concentration of lipid found in the leaf-sheath. The phospholipid compositions of nodes and internodes were similar, comprising mostly phosphatidylcholine (PC) and phosphatidylethanolamine (PE), with linoleic, linolenic and palmitic acids as the predominant component fatty acids. Leaf-sheath tissue contained mainly PE, with equally high amounts of palmitic, linoleic and linolenic acids. β-Sitosterol and cholesterol were the major 4-desmethylsterols of the internode, while β-sitosterol was predominant in the node and leaf-sheath tissues. The growth temperature of segments prior to isolation produced its major effect on the concentration of stigmasterol, which decreased markedly with temperature. The sitosterol/stigmasterol ratio increased significantly as temperature decreased. Stem segments isolated from plants treated with gibberellic acid (GA₃) for 3 weeks, showed a significant reduction in the amounts of 4-desmethyl sterols on a dry wt basis when compared with control segments. However, when stem segments were incubated with GA₃ for 20 hr, no change in 4-desmethylsterol composition or concentration was observed, even though significant growth in response to GA₃ occurred.     

Sterols in Laodelphax striatellus with special reference to the intracellular yeastlike symbiotes as a sterol source

Sterols were analysed to investigate the sterol source in Laodelphax striatellus and three other rice plant-sucking homopterous insects. In L. striatellus, cholesterol, 24-methylenecholesterol and β-sitosterol were detected. The host plant (rice) contained campesterol, stigmasterol and β-sitosterol. From the honeydew excreted by L. striatellus, cholesterol, β-sitosterol and negligible amounts of campesterol were recovered. Laodelphax striatellus possesses yeastlike symbiotes which can be destroyed by high temperature. Fifth instar nymphs, which have been exposed to 35°C for 3 days in their 1 st instar, showed lower cholesterol concentration and markedly reduced amounts of 24-methylenecholesterol. From the results it is concluded that L. striatellus has two sterol sources: one from the host plant and the other from the yeastlike symbiotes which appear to provide 24-methylenecholesterol.     

Circulating sterols as predictors of early allograft dysfunction and clinical outcome in patients undergoing liver transplantation

Introduction

Sensitive and specific assessment of the hepatic graft metabolism after liver transplantation (LTX) is essential for early detection of postoperative dysfunction implying the need for consecutive therapeutic interventions.

Objectives

Here, we assessed circulating liver metabolites of the cholesterol pathway, amino acids and acylcarnitines and evaluated their predictive value on early allograft dysfunction (EAD) and clinical outcome in the context of LTX.

Methods

The metabolites were quantified in the plasma of 40 liver graft recipients one day pre- and 10 days post-LTX by liquid chromatography/tandem mass spectrometry (LC–MS/MS). Plant sterols as well as cholesterol and its precursors were determined in the free and esterified form; lanosterol in the free form only. Metabolites and esterification ratios were compared to the model for early allograft function scoring (MEAF) which is calculated at day 3 post-LTX from routine parameters defining EAD.

Results

The hepatic esterification ratio of all sterols, but not amino acids and acylcarnitine concentrations, showed substantial metabolic disturbances post-LTX and correlated to the MEAF. In ROC analysis, the low esterification ratio of β-sitosterol and stigmasterol from day 1 and of the other sterols from day 3 were predictive for a high MEAF, i.e. EAD. Additionally, the ratio of esterified β-sitosterol and free lanosterol were predictive for all days and the esterification ratio of the other sterols at day 3 or 4 post-LTX for 3-month mortality.

Conclusion

Low ratios of circulating esterified sterols are associated with a high risk of EAD and impaired clinical outcome in the early postoperative phase following LTX.

     

Patterns of lipid biomarkers in the fruit of Bupleurum L. growing wild in Turkey

Four Bupleurum species, representing two sections, were collected from their natural habitats and the fatty acid and sterol compositions and α-tocopherol contents of their fruit oils examined. The major fatty acids were oleic, linoleic and palmitic acids in all species (41.13–79.92%, 11.07–23.21% and 3.52–7.60%, respectively). Δ-5-Avenasterol, stigmasterol, β-sitosterol, Δ-5,24-stigmastadienol and campesterol exhibited in significant concentrations in all species (9.63–48.79%, 15.42–33.44%, 16.11–25.35%, 12.67–22.97% and 3.10–4.86%, respectively). Higher α-tocopherol concentrations were quantified in both species from section Eubupleura (2.55–2.70 mg/100 g) than in section Perfoliata. Significant correlations were found among the fatty acid and phytosterol quantities observed in the Bupleurum accessions. The results indicated that these species have potential as alternative raw materials and nutraceutical sources through the selection of high yield cultivars and agricultural production.     

Functional expression of Arabidopsis thaliana sterol glycosyltransferase from stably transformed Drosophila melanogaster S2 cells

Arabidopsis thaliana sterol glycosyltransferase (SGT), UGT80A2, was expressed from stably transformed Drosophila melanogaster Schneider 2 (S2) cells. Recombinant SGT was detected in both intracellular and extracellular fractions with a molecular mass of approximately 76 kDa. Secreted recombinant SGT accounted for approximately 60% of the total recombinant SGT production. Recombinant SGT in the extracellular fractions was purified to homogeneity using a simple one-step Ni-NTA affinity fractionation. Radiometrical assay using uridine diphospho-d-[U-¹⁴C]glucose (UDP-¹⁴C-glucose) as a sugar donor and sterols, β-sitosterol and stigmasterol, as sugar acceptors showed that the purified recombinant SGT contained UDP-glycosyltransferase activity and could attach ¹⁴C-glucose to β-sitosterol and stigmasterol. Recombinant SGT contained higher catalytic activity with β-sitosterol, which was similar to the recombinant SGT produced by a bacterial expression system. The transfer of ¹⁴C-glucose by recombinant SGT was further determined by gas chromatography-mass spectrometry (GC-MS) analysis of cellulase-treated ¹⁴C-glucosetransferred β-sitosterol and stigmasterol reactants.     

Characterizing components of the Saw Palmetto Berry Extract (SPBE) on prostate cancer cell growth and traction

Saw Palmetto Berry Extract (SPBE) is applied for prostate health and treatment of urinary tract infections, nonbacterial prostitis and Benign Prostatic Hyperplasia (BPH) in man. An assumption is that SPBE affects tumor cell progression and migration in breast and prostate tissue. In this work, DU-145 cells were used to demonstrate that SPBE and its sterol components, β-sitosterol and stigmasterol, inhibit prostate cancer growth by increasing p53 protein expression and also inhibit carcinoma development by decreasing p21 and p27 protein expression. In the presence of cholesterol, these features are not only reversed but increased significantly. The results show for the first time the potential of SPBE, β-sitosterol and stigmasterol as potential anti-tumor agents.Since the protein p53 is also regarded as nuclear matrix protein facilitating actin cytoskeletal binding, 2D tractions were measured. The cell adhesion strength in the presence of SPBE, β-sitosterol and cholesterol and the observation was that the increase in p53 expression triggered an increase in the intracellular force generation. The results suggest a dual function of p53 in cells.     

Post-pollination hybridization barriers in <Emphasis Type="Italic">Nicotiana</Emphasis> section <Emphasis Type="Italic">Alatae</Emphasis>

Nicotiana section Alatae contains eight species with variable flower sizes and morphologies. Section members readily hybridize in the glasshouse, but no hybrids have been observed in natural sympatric and parapatric populations. To investigate interspecific crossing relationships with respect to mechanisms preventing hybridization, all members of section Alatae were intercrossed in a complete diallel. We found positive correlation between the pistil length of the pollen donor and interspecific seed set relative to the conspecific cross. Pollen tube growth rate and pollen donor pistil length were positively correlated as well. Furthermore, pollen from short-pistil members of section Alatae could only grow a maximum distance proportional to, but greater than, their own pistil lengths. Our results show that pollen tube growth capacity (i.e., rate and distance), provides a hybridization barrier in long-pistil species × short-pistil species crosses. We also found another hybridization barrier not specifically related to pollen tube growth capacity in short-pistil species × long-pistil species. Taken together, these barriers can generally be described by a ‘pistil-length mismatch’ rule; in section Alatae, pollen has the most success fertilizing ovules from species with pistil lengths similar to their own. This rule could contribute to hybridization barriers in Section Alatae because the species display dramatically different pistil lengths. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.